Combinatorial On‐Chip Study of Miniaturized 3D Porous Scaffolds Using a Patterned Superhydrophobic Platform

Abstract

One of the main challenges in tissue engineering (TE) is to obtain optimized products, combining biomaterials, cells and soluble factors able to stimulate tissue regeneration. Multiple combinations may be considered by changing the conditions among these three factors. The unpredictable response of each combination requires time-consuming tests. High-throughput methodologies have been proposed to master such complex analyses in TE. Usually, these tests are performed using cells cultured into 2D biomaterials or by dispensing arrays of cell-loaded hydrogels. For the first time an on-chip combinatorial study of 3D miniaturized porous scaffolds is proposed, using a patterned bioinspired superhydrophobic platform. Arrays of biomaterials are dispensed and processed in situ as porous scaffolds with distinct composition, surface characteristics, porosity/pore size, and mechanical properties. On-chip porosity, pore size, and mechanical properties of scaffolds based on chitosan and alginate are assessed by adapting microcomputed tomography equipment and a dynamic mechanical analyzer, as well as cell response after 24 hours. The interactions between cell types of two distinct origins-osteoblast-like and fibroblasts-and the scaffolds modified with fibronectin are studied and validated by comparison with conventional destructive methods (dsDNA quantification and MTS tests). Physical and biological on-chip analyses are coherent with the conventional measures, and conclusions about the most favorable conditions for each cell type are taken.

Towards the design of 3D multiscale instructive tissue engineering constructs: Current approaches and trends

“…High-content screening of 3D scaffolds properties and features promises a huge contribute in deciphering which cues, materials and methods will be more effective for the regeneration of each tissue. (Hook, et al, 2010;Simon and Lin-Gibson, 2011;Oliveira, et al, 2013a; 2014a)…”

In Vivo High-Content Evaluation of Three-Dimensional Scaffolds Biocompatibility

“…We previously proved that these Chi concentrations are adequate for the preparation of porous scaffolds on such chips. 21 Nine scaffolds of each concentration were prepared in the chips (Fig. 2D).…”

“…[18][19][20] Similar platforms were used in vitro for the study of protein-cell interactions in two-dimensional (2D) environment, cell-laden three-dimensional (3D) hydrogels, and 3D scaffolds. [19][20][21] We hypothesize that the miniaturized size of the patterned biomaterials and the gap maintained between them would be sufficient to observe distinct inflammatory cells recruitment while allowing for isolated responses in each spot. Moreover, the low cell adhesion reported in superhydrophobic surfaces would improve the independency between spots.…”

Designing biomaterials for tissue engineering based on the deconstruction of the native cellular environment

“…Multiple solutions of organising biomaterials to be tested have been proposed for such analysis, usually in the form of arrays or gradients. We developed a platform based on superhydrophobic substrates exhibiting patterned wettable spots to support 3D biomaterials for both in-vitro [8,9] and in-vivo [10] high-throughput screening. High-throughput methodologies permit to work under reduced a priori assumptions because any possible combination will be considered as equally valid as the others before the testing.…”